WO2007147454A1

WO2007147454A1 - Chemical synthesis of cu-64asp (copper-64asparaginase; [64cu]asparaginase) for pet scans and radiotheraphy

Info

Publication number: WO2007147454A1
Application number: PCT/EP2007/001317
Authority: WO
Inventors: Gianluca Valentini; Paola Panichelli; Domenico Martini
Original assignee: A.C.O.M. Advanced Center Oncology Macerata S.P.A.
Priority date: 2006-06-23
Filing date: 2007-02-15
Publication date: 2007-12-27
Also published as: ITMC20060080A1

Abstract

This invention regards a new molecule: the Cu-64ASP (copper- 64asparaginase; [64CU]asparaginase), which, used in the PET diagnostic methodology, allows a double in “vivo” use: the diagnostic and radiotherapeutical ones.

Description

Title: "Chemical synthesis of Cu-64Asp (copper-64asparaginase; [⁶⁴Cu]asparaginase) for PET scans and radiotherapy".

§§§ This invention regards a new molecule: the Cu-64ASP (copper- 64asparaginase; [⁶⁴CU] asparaginase), which, used in the PET diagnostic methodology, allows a double in "vivo" use: the diagnostic and radio- therapeutical ones.

A PET (Position Emission Tomography) scan is a diagnostic methodology which permits the study of the metabolism of each examined organ, allowing to obtain an early diagnosis of the different pathologies.

In the last few years, such methodology has had a great expansion in the USA and, now, PET's diagnostic centres are growing throughout Europe, especially in oncology. In that field, PET scans introduce two clinical parameters, such as early diagnosis and therapy optimization, which modify life expectancy of patients and improve the management of their illness.

Other examples of its fields of application, such as neurology, cardiology and rheumatology, are gaining greater importance. In a time when we continually notice an increment in the average lifetime, the role PET scan will acquire in the study of the diseases linked to aging, such as Parkinson and Alzheimer, as well as in the study of molecules that would give the possibility to precociously diagnose the onset of acute cardiac events that nowadays represent one of the main causes of death, will be very important. To carry out a PET scan, the patient is injected with a radiodrug, which is later followed through the human body using specific machinery called PET Tomographs.

Radiodrugs consist of two essential parts, which are a radioisotope (that emits beta radiations) and a molecule that will be bound with the radioisotope and is the metabolic substrate of the PET scan. Radioisotopes are produced through a tool called Cyclotron and are bound with the molecule under study, through specific chemical synthesis methods.

The radioisotope commonly used today is the 18-Fluorine, which has a two hours half-life and has chemical characteristics suitable to be easily bound, in its liquid form, to various molecules.

The molecule most commonly used today is the 18-Fluorine-deoxyglucose (FDG), which, in human beings, allows the identification of the locations that use a greater amount of glucose, compared to the normal metabolic standards and that, therefore, could have a carcinogenic nature. The limitation rising from such application is a non-sufficient specificity in distinguishing tumoral sites from those, which are possible site of infection. In this sense, other molecules, such as Fluorine-choline or Fluorine- thymidine, which increase diagnostic specificity, especially in tumoral forms as prostate cancer or brain and lung cancer, have been experimented. Another field of great development in the PET diagnostic methodology is the development of radiotherapy treatment plans, which allow the optimization of the therapeutical effect. In this sense, in this field research, there are various development phases, which, starting from a demonstrated benefit of FDG, get the most recent experiences made with the ⁶⁴Cu-ATSM molecule that permits to obtain a map of the distribution of oxygen inside the tumoral mass. Such molecule, synthesized in the USA, modifies the treatments plans in its ability to reduce the area of intervention as well as in its ability to exclude the areas that don't present oxygen and that, therefore, don't present any possibility to answer to radiotherapy. There are experiments like that, also in molecules labeled with 18-Fluorine, such as the F-MISO, but the half-life characteristics of Copper-64, lasting about 12 hours, today, make it a radioisotope of great perspective interest of the field. The invention herein regards the experimentation and the synthesis of a new molecule, Copper-64asparaginase, where copper is able to emit short-range electrons, having a direct anti-tumoral effect.

Furthermore, the synthesis of the two components creates the molecule subject of this application, which also has the characteristic of having a double potential of its in "vivo" use, with reference to the diagnostic and the radio-therapeutical treatments.

Copper-64 comes from a Nickel-64 electrodeposition ([64]Ni) on a solid target made of a gold disc 24mm of diameter and 2mm thick; to the 30mg of [64]Ni must be added 3ml of nitric acid 99,99999% pure and 3ml of H₂O Millipore; then, it must be warmed up until the solution evaporates completely, leaving behind a green residue. Then, 2ml of sulphuric acid 99,9999% pure and 2ml Of H₂O millipore are added drop by drop; Then it's warmed up until it evaporates completely, and then 3,5 ml of H₂O millipore are added and warmed up again. Its final volume is 3 ml, it is cooled at Room Temperature and than 28% pure ammonia is added, until it changes color. Later, 0,1 g of ammonium sulphate 99,9999% pure is added. Retrieval of Copper-64 (⁶⁴Cu) obtained by the nuclear reaction ⁶⁴Ni(p,n)⁶⁴Cu, through the 14 Mev IBA cyclotron: the solid target is treated under heat for 20 minutes, with 6 ml of HCL 6 M, to dissolve the electrodeposition. The obtained solution is poured into a column parceled with the AG 1x8 resin, produced by Biorad, conditioned with 10-12 ml of HCL 99,999% pure and 15 ml of HCL are added to the solution. Then, through a 0,5 M solution of HCL, [64]Ni is retrieved and Copper-64 chloride (⁶⁴CuCl₂) is flowed. We withdraw 10 mCi Of ⁶⁴CuCl₂, evaporate it under a reduced pressure and we add 100 μl of pH 5.5 acetate buffer and a solution of Asparaginase and L-Methionine sulfoximine melted in 300 μl of pH 5.5 acetate buffer. The solution goes through a 0,22 um filter and is diluted with a saline solution. An alternative synthesis is to use Asparaginase-polyethylene glycol, rather than Asparaginase.

Claims

C L A I M S

1. Molecule of Copper-64asparaginase, which synthesis of the two components is characterized by having a double potential of in "vivo" use, with reference to diagnostic and radio-therapeutical treatments and where copper is able to emit short-range electrons, which have a direct anti-tumoral effect;

2. Synthesis process of complexes of copper-64 with Asparaginase, pursuant to claim number 1, characterized by the presence of L-Methionine sulfoximine with a coordination of the Asparaginase macromolecule to the metal, through a reaction at room temperature.

3. The molecule of Copper-64 Asparaginase-poly ethylene-glycol, which synthesis of its two components is characterized by having a double potential of in "vivo" use, with reference to diagnostic and radio-therapeutical treatments and where copper is able to emit short-range electrons, which have a direct anti-tumoral effect;

4. Synthesis process of complexes of copper-64 with Asparaginase- poly ethylene-glycol, pursuant to claim number 3, characterized by the presence of L-Methionine sulfoximine with coordination of the 64Asparaginase-polyethylene-glycol macromolecule to the metal, through a reaction at room temperature.

5. The Copper-64asparaginase molecule, pursuant to claims number 1 and 2, characterized by the coordination of asparaginase to the metal, through a reaction at room temperature;

6. The Copper-64Asparaginase-polyethylene-glycol molecule, pursuant to claims number 3 and 4, characterized by the coordination of Asparaginase- polyethylene-glycol to the metal, through a reaction at room temperature;

7. The Copper-64Asparaginase molecule, pursuant to claims number 1, 2 and 5, characterized by the fact that it can be administered through general parenteral route in the form of aqueous or oily solutions or suspended in appropriate dispersing agents, in the form of lyophilized products to dispel at the moment of the administration;

8. The Copper-64Asparaginase-polyethylene-glycol molecule, pursuant to claims number 3, 4 and 6, characterized by the fact that it can be administered through general parenteral route in the form of aqueous or oily solutions, or suspended in appropriate dispersing agents, even in the form of lyophilized products, to dispel at the moment of the administration;

9. The Copper-64Asparaginase molecule, pursuant to claims number 1, 2, 5 and 7, characterized by the fact that it can be administered through general oral route in the form of tablets, soft or hard gelatin oily or percolated capsules, sugar-coated pills, dispersing powder, suspensions or emulsions;

10. The Copper-64Asparaginase-poly ethylene-glycol molecule, pursuant to claims number 3, 4, 6 and 8, characterized by the fact that it can be administered through general oral route in the form of tablets, soft or hard gelatin oily or percolated capsules, sugar-coated pills, dispersing powders, suspensions or emulsions;

11. The Copper-64 Asparaginase molecule, pursuant to claims number 1, 2, 5, 7 and 9, characterized by the fact that it can be administered through topical or transdermal route in the appropriate forms and in vehicles or devices suited for the administration of the active principle at the primary and/or secondary site of the tumor;

12. The Copper-64Asparaginase-polyethylene-glycol molecule, pursuant to claims number 3, 4, 6 and 8, characterized by the fact that it can be administered through topical or transdermal route in the appropriate forms and in vehicles or devices suited for the administration of the active principle at the primary and/or secondary site of the tumor;

13. The Copper-64 Asparaginase molecule, pursuant to all of the above claims and according to what is described and explained for specified purposes.